New categorization of human vascular endothelial cells by pro- vs anti-proliferative phenotypes

Figure 3 Regulator of G-protein signaling 5 is involved in the phenotype conversion of vascular endothelial cells. A: qRT-PCR results of RGS5 message expression were shown with normalization by GAPDH. The horizontal axis was demonstrated as logarithm; B: Immunostaining of indicated cells by using an anti-RGS5 antibody (red) with DAPI counterstaining (blue); C: Western blotting of RGS5 protein; D: Clinical specimens of the artery were subjected to immunostaining studies by using an anti-RGS5 antibody (red) and an anti-human PECAM1 antibody (green) with nuclear counterstaining with DAPI (blue); E-G: RGS5 knockdown experiments. An expression vector for shRNA against RGS5 (Sh-RGS5) or control RNA (CTL RNA) were transfected into EPC1dEC[P9] (E) or HUVEC (F). The results of VSMC co-cultured experiments (E and F) along with qRT-PCR (RGS5/GAPDH) (G) and Western blotting (H) regarding HUVEC were shown. GAPDH: Glyceraldehyde-phosphate dehydrogenase; VSMC: Vascular smooth muscle cell; VEC: Vascular endothelial cell; iPSC: Induced pluripotent stem cell; PCR: Polymerase chain reaction; RGS5: Regulator of G-protein signaling 5; HUVEC: Human umbilical vein endothelial cell; HAEC: Human adult aortic endothelial cells; HMVEC: Human neonatal dermal microvascular endothelial cells.